Articles
CLONING OF GENES CONFERRING ALLERGENIC PROTEINS AND SALT TOLERANCE BY SCREENING A FULL LENGTH CDNA LIBRARY IN SOYBEAN
Article number
929_18
Pages
129 – 134
Language
English
Abstract
To obtain genes conferring allergenic proteins and salt tolerance of vegetable soybean (Glycine max (L.) Merr.), a full-length cDNA library from the developing seeds of NN99-10 was constructed and characterized.
Gel-electrophoresis results showed that most of the cDNA inserts ranged from 0.40 to 2.0 kb, with an average size c. 800 bp.
Randomly-picked clones showed that the recombination rate was >99%. This indicated that the cDNA library constructed was a full-length library with high quality.
After blue/white selection followed by sequencing of the positive cDNA clones from the constructed library, a total of 642 expressed sequence tags (ESTs) were identified, 468 of which contained full-length coding regions.
Based on bioinformatic analysis of the sequences, a total of 395 full-length cDNA clones were homologous to various known genes in soybean and other species.
The remaining 73 full-length cDNA clones showed little homology to known sequences, which might be considered novel.
The identified 395 full-length cDNA clones fell into 8 functional categories, including modifications and precursors of protein (26.31%), protein synthesis (22.24%), and defense proteins (22.20%). A total of eleven genes of agronomic importance were identified, among which eight related to seed quality (four genes conferring allergenic proteins) and three conferring salt tolerance.
Gel-electrophoresis results showed that most of the cDNA inserts ranged from 0.40 to 2.0 kb, with an average size c. 800 bp.
Randomly-picked clones showed that the recombination rate was >99%. This indicated that the cDNA library constructed was a full-length library with high quality.
After blue/white selection followed by sequencing of the positive cDNA clones from the constructed library, a total of 642 expressed sequence tags (ESTs) were identified, 468 of which contained full-length coding regions.
Based on bioinformatic analysis of the sequences, a total of 395 full-length cDNA clones were homologous to various known genes in soybean and other species.
The remaining 73 full-length cDNA clones showed little homology to known sequences, which might be considered novel.
The identified 395 full-length cDNA clones fell into 8 functional categories, including modifications and precursors of protein (26.31%), protein synthesis (22.24%), and defense proteins (22.20%). A total of eleven genes of agronomic importance were identified, among which eight related to seed quality (four genes conferring allergenic proteins) and three conferring salt tolerance.
Authors
Yuelin Zhu, Lifei Yang, Junyi Gai
Keywords
Glycine max, gene cloning, full-length cDNA library, allergenic protein, salt tolerance
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